Target (protein/gene name): Protein Phosphatase 2C *NCBI Gene # or RefSeq#: ABO31335.1 GI:129716418 *Protein ID (NP or XP #) or Wolbachia#:
*Organism (including strain): Toxoplasma Gondii
Etiologic Risk Group (see link below):
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): Toxoplasma Gondii is a eukaryotic parasitic protozoan that causes toxoplasmosis. This organism is found worldwide and capable of infecting all warm-blooded animals. As one of the most common human parasites, T. Gondii has been predicted to have infected up to a third of the global population. Although the parasite is found worldwide, the disease itself (toxoplasmosis) only causes mild flu-like symptoms. However in infants and HIV/AIDS patients, this infection cause serious and potentially fatal illness. Infection of this disease usually occurs due to consumption of raw/undercooked meat, ingesting soil/vegetables/water contaminated with cat feces, or through transmission from mother to fetus during pregnancy if mother is infected.
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): ORIGINAL:
maslrvlalaclplfavlhadaktllgkvkratgmgvgegpsvakkpkytatvpgftpps gdqrmnefmavdtsgefmrhlyieegrtvcasatsrnrrptsesphsddvvvvegmlrgr petrvhamfdgfqgrhsamwlaqnvmnylndlrdvneeeitrqfermdgdlraanlpggs saliifvryekkptearvvgrqivpegakeftsvaealggplmpvvamnfrrdpraakgi ytihvaslgnsrcvlksgrtaihlstphtasshkerhrvqaaggvfttvngelllggvvp mtrafgsfdfkkggqgklqqdlvsavpdvttffaypgddivagtagafahfrshaaiaaa ialypvspetvldaakamvvnakrrkvtknistfvrhlpesrtrsqkmlegtsgengeed fsidrtneltqalqagffsfllyyp
*length of your protein in Amino Acids- ORIGINAL: 445 Cut Sequence: 400
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParamwebsite: 43297.3 Da Molar Extinction coefficient of your protein at 280 nm wavelength: TMpred graph Image
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1800361/
Protein Link from Paper:
http://www.ncbi.nlm.nih.gov/protein/?Db=protein&DbFrom=pmc&Cmd=Link&LinkName=pmc_protein&IdsFromResult=1800361
Link to Protein Phosphatase 2C:
http://www.ncbi.nlm.nih.gov/protein/ABO31335.1
Target (protein/gene name): Protein Phosphatase 2C
*NCBI Gene # or RefSeq#: ABO31335.1 GI:129716418
*Protein ID (NP or XP #) or Wolbachia#:
*Organism (including strain): Toxoplasma Gondii
Etiologic Risk Group (see link below):
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): Toxoplasma Gondii is a eukaryotic parasitic protozoan that causes toxoplasmosis. This organism is found worldwide and capable of infecting all warm-blooded animals. As one of the most common human parasites, T. Gondii has been predicted to have infected up to a third of the global population. Although the parasite is found worldwide, the disease itself (toxoplasmosis) only causes mild flu-like symptoms. However in infants and HIV/AIDS patients, this infection cause serious and potentially fatal illness. Infection of this disease usually occurs due to consumption of raw/undercooked meat, ingesting soil/vegetables/water contaminated with cat feces, or through transmission from mother to fetus during pregnancy if mother is infected.
Essentiality of this protein: Essential
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1800361/
Complex of proteins?:
Druggable Target:
*EC#: 3.1.3.16
Link to BRENDA EC# page: http://www.brenda-enzymes.org/php/result_flat.php4?ecno=3.1.3.16
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
-- List cost and quantity of substrate reagents and supplier
Structure Available (PDB): http://www.rcsb.org/pdb/explore/explore.do?structureId=3D8K
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells):
Purification Method:
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
ORIGINAL:
maslrvlalaclplfavlhadaktllgkvkratgmgvgegpsvakkpkytatvpgftpps gdqrmnefmavdtsgefmrhlyieegrtvcasatsrnrrptsesphsddvvvvegmlrgr petrvhamfdgfqgrhsamwlaqnvmnylndlrdvneeeitrqfermdgdlraanlpggs saliifvryekkptearvvgrqivpegakeftsvaealggplmpvvamnfrrdpraakgi ytihvaslgnsrcvlksgrtaihlstphtasshkerhrvqaaggvfttvngelllggvvp mtrafgsfdfkkggqgklqqdlvsavpdvttffaypgddivagtagafahfrshaaiaaa ialypvspetvldaakamvvnakrrkvtknistfvrhlpesrtrsqkmlegtsgengeed fsidrtneltqalqagffsfllyyp
"CUT DOWN" SEQUENCE (FROM RESEARCH PAPER):
madaktllgkvkratgmgvgegpsvakkpkytatvpgftpps gdqrmnefmavdtsgefmrhlyieegrtvcasatsrnrrptsesphsddvvvvegmlrgr petrvhamfdgfqgrhsamwlaqnvmnylndlrdvneeeitrqfermdgdlraanlpggs saliifvryekkptearvvgrqivpegakeftsvaealggplmpvvamnfrrdpraakgi ytihvaslgnsrcvlksgrtaihlstphtasshkerhrvqaaggvfttvngelllggvvp mtrafgsfdfkkggqgklqqdlvsavpdvttffaypgddivagtagafahfrshaaiaaa ialypvspetvldaakamvvnakrrkvtknistfvrhlpesrtrsqkmlegtsgenge
*length of your protein in Amino Acids- ORIGINAL: 445
Cut Sequence: 400
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParamwebsite: 43297.3 Da
Molar Extinction coefficient of your protein at 280 nm wavelength:
TMpred graph Image
*CDS Gene Sequence (paste as text only):
ATGGCGGATGCTAAAACCCTGCTCGGTAAAGTTAAACGTGCGACCGGTATGGGTGTTGGTGAAGGTCCGTCTGT
TGCGAAAAAACCGAAATACACCGCGACCGTTCCTGGCTTTACTCCGCCGTCTGGTGACCAGCGTATGAACGAGT
TCATGGCAGTTGACACCTCTGGCGAATTCATGCGCCACCTGTACATCGAGGAAGGTCGTACCGTTTGCGCGTCT
GCGACTTCTCGTAATCGTCGCCCGACCTCTGAATCTCCGCACTCTGACGATGTTGTTGTGGTTGAGGGTATGCT
GCGTGGTCGTCCGGAAACCCGTGTTCACGCGATGTTCGACGGTTTCCAGGGTCGTCACTCTGCGATGTGGCTGG
CGCAGAACGTTATGAACTACCTGAACGACCTGCGCGACGTCAACGAGGAGGAAATTACCCGTCAGTTTGAACGC
ATGGACGGTGATCTCCGTGCAGCGAACCTGCCGGGTGGCTCTAGCGCCCTGATCATCTTTGTTCGCTACGAAAA
GAAACCAACCGAAGCGCGTGTAGTTGGCCGTCAGATCGTCCCGGAGGGTGCGAAGGAATTCACTTCTGTGGCGG
AAGCGCTGGGCGGTCCACTGATGCCTGTCGTTGCGATGAACTTCCGTCGTGACCCGCGTGCTGCGAAAGGTATC
TACACCATCCACGTAGCGTCTCTGGGTAACTCTCGTTGTGTACTGAAATCTGGCCGTACCGCGATCCACCTCTC
TACCCCGCACACTGCCTCTTCTCACAAAGAACGCCACCGCGTACAAGCCGCTGGCGGTGTCTTTACGACGGTAA
ATGGTGAACTGCTGCTGGGTGGTGTTGTCCCGATGACCCGTGCGTTCGGTTCTTTCGACTTCAAAAAAGGCGGT
CAGGGTAAACTGCAGCAGGACCTCGTTTCTGCGGTTCCTGATGTTACCACCTTCTTCGCGTACCCAGGCGATGA
CATCGTTGCAGGTACTGCGGGTGCTTTCGCGCACTTTCGTTCTCATGCCGCAATCGCTGCGGCTATTGCCCTGT
ATCCGGTTTCTCCTGAGACTGTTCTGGACGCCGCAAAGGCTATGGTTGTTAACGCGAAACGTCGCAAAGTTACC
AAAAACATCTCTACGTTCGTTCGTCACCTCCCGGAATCTCGTACCCGTTCCCAGAAAATGCTGGAAGGTACTTC
CGGTGAAAACGGCGAATAA
*GC% Content for gene: 55.36%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
TACTTCCAATCCATGGCGGATGCTAAAACCCTGCTCGGTAAAGTTAAACGTGCGACCGGTATGGGTGTTGGTGAAGGTCCGTC
TGTTGCGAAAAAACCGAAATACACCGCGACCGTTCCTGGCTTTACTCCGCCGTCTGGTGACCAGCGTATGAACGAGTTCATGG
CAGTTGACACCTCTGGCGAATTCATGCGCCACCTGTACATCGAGGAAGGTCGTACCGTTTGCGCGTCTGCGACTTCTCGTAAT
CGTCGCCCGACCTCTGAATCTCCGCACTCTGACGATGTTGTTGTGGTTGAGGGTATGCTGCGTGGTCGTCCGGAAACCCGTGT
TCACGCGATGTTCGACGGTTTCCAGGGTCGTCACTCTGCGATGTGGCTGGCGCAGAACGTTATGAACTACCTGAACGACCTGC
GCGACGTCAACGAGGAGGAAATTACCCGTCAGTTTGAACGCATGGACGGTGATCTCCGTGCAGCGAACCTGCCGGGTGGCTCT
AGCGCCCTGATCATCTTTGTTCGCTACGAAAAGAAACCAACCGAAGCGCGTGTAGTTGGCCGTCAGATCGTCCCGGAGGGTGC
GAAGGAATTCACTTCTGTGGCGGAAGCGCTGGGCGGTCCACTGATGCCTGTCGTTGCGATGAACTTCCGTCGTGACCCGCGTG
CTGCGAAAGGTATCTACACCATCCACGTAGCGTCTCTGGGTAACTCTCGTTGTGTACTGAAATCTGGCCGTACCGCGATCCAC
CTCTCTACCCCGCACACTGCCTCTTCTCACAAAGAACGCCACCGCGTACAAGCCGCTGGCGGTGTCTTTACGACGGTAAATGG
TGAACTGCTGCTGGGTGGTGTTGTCCCGATGACCCGTGCGTTCGGTTCTTTCGACTTCAAAAAAGGCGGTCAGGGTAAACTGC
AGCAGGACCTCGTTTCTGCGGTTCCTGATGTTACCACCTTCTTCGCGTACCCAGGCGATGACATCGTTGCAGGTACTGCGGGT
GCTTTCGCGCACTTTCGTTCTCATGCCGCAATCGCTGCGGCTATTGCCCTGTATCCGGTTTCTCCTGAGACTGTTCTGGACGC
CGCAAAGGCTATGGTTGTTAACGCGAAACGTCGCAAAGTTACCAAAAACATCTCTACGTTCGTTCGTCACCTCCCGGAATCTC
GTACCCGTTCCCAGAAAATGCTGGAAGGTACTTCCGGTGAAAACGGCGAATAACAGTAAAGGTGGATA
*GC% Content for gene (codon optimized): 55.04%
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
Forward: TACTTCCAATCCATGGCGGATGCTAAAACC
Reverse: TATCCACCTTTACTGTTATTCGCCGTTTTCACC
The oligonucleotide primer sequences:
1 ATGGCGGATGCTAAAACCCTGCTCGGTAAAGTTAAACGTGCG 42
2 TTTCGCAACAGACGGACCTTCACCAACACCCATACCGGTCGCACGTTTAACTTTACCGAG 60
3 GGTCCGTCTGTTGCGAAAAAACCGAAATACACCGCGACCGTTCCTGGCTTTACTCCGCCG 60
4 AGGTGTCAACTGCCATGAACTCGTTCATACGCTGGTCACCAGACGGCGGAGTAAAGCCAG 60
5 GTTCATGGCAGTTGACACCTCTGGCGAATTCATGCGCCACCTGTACATCGAGGAAGGTCG 60
6 CGGGCGACGATTACGAGAAGTCGCAGACGCGCAAACGGTACGACCTTCCTCGATGTACAG 60
7 TCTCGTAATCGTCGCCCGACCTCTGAATCTCCGCACTCTGACGATGTTGTTGTGGTTGAG 60
8 CGCGTGAACACGGGTTTCCGGACGACCACGCAGCATACCCTCAACCACAACAACATCGTC 60
9 AAACCCGTGTTCACGCGATGTTCGACGGTTTCCAGGGTCGTCACTCTGCGATGTGGCTGG 60
10 TTGACGTCGCGCAGGTCGTTCAGGTAGTTCATAACGTTCTGCGCCAGCCACATCGCAGAG 60
11 ACCTGCGCGACGTCAACGAGGAGGAAATTACCCGTCAGTTTGAACGCATGGACGGTGATC 60
12 TGATCAGGGCGCTAGAGCCACCCGGCAGGTTCGCTGCACGGAGATCACCGTCCATGCGTT 60
13 GCTCTAGCGCCCTGATCATCTTTGTTCGCTACGAAAAGAAACCAACCGAAGCGCGTGTAG 60
14 AAGTGAATTCCTTCGCACCCTCCGGGACGATCTGACGGCCAACTACACGCGCTTCGGTTG 60
15 GGGTGCGAAGGAATTCACTTCTGTGGCGGAAGCGCTGGGCGGTCCACTGATGCCTGTCGT 60
16 ATACCTTTCGCAGCACGCGGGTCACGACGGAAGTTCATCGCAACGACAGGCATCAGTGGA 60
17 GCGTGCTGCGAAAGGTATCTACACCATCCACGTAGCGTCTCTGGGTAACTCTCGTTGTGT 60
18 CGGGGTAGAGAGGTGGATCGCGGTACGGCCAGATTTCAGTACACAACGAGAGTTACCCAG 60
19 GATCCACCTCTCTACCCCGCACACTGCCTCTTCTCACAAAGAACGCCACCGCGTACAAGC 60
20 CCAGCAGCAGTTCACCATTTACCGTCGTAAAGACACCGCCAGCGGCTTGTACGCGGTGGC 60
21 AATGGTGAACTGCTGCTGGGTGGTGTTGTCCCGATGACCCGTGCGTTCGGTTCTTTCGAC 60
22 ACGAGGTCCTGCTGCAGTTTACCCTGACCGCCTTTTTTGAAGTCGAAAGAACCGAACGCA 60
23 CTGCAGCAGGACCTCGTTTCTGCGGTTCCTGATGTTACCACCTTCTTCGCGTACCCAGGC 60
24 GAAAGTGCGCGAAAGCACCCGCAGTACCTGCAACGATGTCATCGCCTGGGTACGCGAAGA 60
25 GTGCTTTCGCGCACTTTCGTTCTCATGCCGCAATCGCTGCGGCTATTGCCCTGTATCCGG 60
26 AACCATAGCCTTTGCGGCGTCCAGAACAGTCTCAGGAGAAACCGGATACAGGGCAATAGC 60
27 GCCGCAAAGGCTATGGTTGTTAACGCGAAACGTCGCAAAGTTACCAAAAACATCTCTACG 60
28 GAACGGGTACGAGATTCCGGGAGGTGACGAACGAACGTAGAGATGTTTTTGGTAACTTTG 60
29 CGGAATCTCGTACCCGTTCCCAGAAAATGCTGGAAGGTACTTCCGGTGAAAACGGCGAAT 60
30 TTATTCGCCGTTTTCACCGG 20